CN112654259A - Functional plant salt and preparation method and application thereof - Google Patents
Functional plant salt and preparation method and application thereof Download PDFInfo
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- CN112654259A CN112654259A CN201980048684.3A CN201980048684A CN112654259A CN 112654259 A CN112654259 A CN 112654259A CN 201980048684 A CN201980048684 A CN 201980048684A CN 112654259 A CN112654259 A CN 112654259A
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/40—Table salts; Dietetic salt substitutes
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
A functional plant salt is prepared by mixing different plant extract powder with salt at a certain ratio, and baking at 800-1400 deg.C for one time or several times to obtain functional plant salt with different colors, spatial structures, physicochemical properties, flavors and uses.
Description
The invention relates to the technical field of food and health food, in particular to functional plant salt and a preparation method and application thereof, and more particularly relates to non-bamboo plant salt and a preparation method and application thereof.
The plant salt is a novel edible salt which is extracted from plant organisms, is in accordance with low sodium salt and multiple mineral trace element groups by pure physical processes and technologies, is in accordance with the proportion of various elements in accordance with the requirements of human bodies, and has certain promotion effects on trace element balance, function regulation, metabolism and health of human bodies.
The plant salt has the main characteristics that: plants are taken as main sources; ② natural low sodium salt and natural balanced salt; ③ the fertilizer is rich in various organic nutrient substances and bioactive components; fourthly, no chemical synthetic substance is added in the processing and production process; it has certain auxiliary treatment and regulation effect on various diseases and related complications caused by mineral element group deficiency or imbalance. For example, it has definite curative effect on diseases and complications such as sodium-sensitive hypertension, type 2 diabetes, hyperlipemia, cardiovascular diseases, tumor, organism immune dysfunction and the like.
The Plant-Salt concept was first proposed by professor panhanjie, the institute of preventive medicine, hong Kong, and was continuously perfected and developed.
Professor panhanjie points out that the efficacy of plant salt has a very close relationship with the activities of seed selection of raw material plants, cultivation of soil, irrigation water sources and the like. The plant salt engineering is just started in China, and has a very wide prospect.
The Panhanjie professor emphasizes that the biological activity of the plant salt depends on the selection of raw plant varieties, variety combinations, the amount of inorganic salt varieties contained in the raw plant varieties, the content and proportion relationship of elements, plant-related biological active ingredients and other factors; the plant salt is mainly used for people who need low-sodium diet and people who suffer from body dysfunction, function reduction and function damage caused by imbalance or lack of body trace elements.
Disclosure of Invention
The invention aims to provide a functional plant salt and a preparation method thereof.
In the invention, the functional plant salt is non-bamboo plant salt, which is a product prepared by taking a non-bamboo extract as a plant raw material.
In a first aspect of the present invention, there is provided a functional plant salt prepared by: one or more plant extract powders are evenly mixed with fine salt according to the addition amount of 1-40 percent and baked at the high temperature of 800-1400 ℃ for one time or several times to obtain the functional plant salt.
In another preferred embodiment, the plant extract is an extract of a substance selected from the group consisting of: plum, pine, tea, lotus, chrysanthemum, pomegranate, Chinese flowering crabapple, edible fungus and algae.
In another preferred example, the plant extract is a plum extract and/or a pine extract.
In another preferred embodiment, the plum extract is selected from the group consisting of: plum extract, plum leaf extract, plum branch extract, plum root extract, or a combination thereof; and/or
The pine extract is selected from the group consisting of: pine needle extract, pine bark extract, pine pollen extract, or a combination thereof.
In another preferred embodiment, the fine salt is selected from the group consisting of: sea salt, lake salt, well salt and mineral salt.
In another preferred embodiment, the fine salt is crude sea salt.
In another preferred example, the functional plant salt is pine salt and/or plum salt.
In another preferred embodiment, the plant extract further comprises a bamboo extract.
In another preferred embodiment, the fine salt is common salt.
In another preferred embodiment, the functional plant salt has a Δ L value of 28 to 60, preferably 30 to 55, more preferably 32 to 50.
In another preferred embodiment, the functional plant salt has a Δ L value of 40 to 60, preferably 42 to 55.
In a second aspect of the present invention, there is provided a method for preparing a functional plant salt according to the first aspect of the present invention, comprising the steps of: one or more plant extract powders are evenly mixed with fine salt according to the addition amount of 1-40 percent and baked at the high temperature of 800-1400 ℃ for one time or several times to obtain the functional plant salt.
In another preferred embodiment, the cooking is performed in a mechanized oven.
In another preferred example, the roasting is performed in a medium frequency smelting furnace.
In a third aspect of the invention, there is provided a food or pharmaceutical composition comprising one or more functional plant salts according to the first aspect of the invention.
In a fourth aspect of the invention, there is provided a use of the functional plant salt of the first aspect of the invention for the preparation of a substance selected from the group consisting of: pharmaceutical adjuvants, health food/beverage, functional food/beverage, table salt, cooking salt, special-purpose cosmetics or personal care products.
In a fifth aspect of the invention, there is provided a method of inhibiting transaminase activity, comprising the steps of: administering to a patient in need thereof one or more inhibiting effective amount of a functional plant salt according to the first aspect of the invention.
In another preferred embodiment, the transaminase is selected from the group consisting of: aspartate Aminotransferase (ALT), glutamate Aminotransferase (AST).
It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.
FIG. 1 is a photograph of the pine salt obtained in example 4.
FIG. 2 is a photograph of the bamboo salt obtained in example 5.
FIG. 3 is a photograph of a plum salt obtained in example 6.
FIG. 4 is a photograph of the pine salt obtained in example 7.
FIG. 5 is a photograph of the bamboo salt obtained in example 8.
FIG. 6 is a photograph of a plum salt obtained in example 9.
FIG. 7 is a photograph of plant salt obtained at different ratios and different process parameters.
Figure 8 is a micrograph of different salts.
Fig. 9 is SEM results for different salts.
Figure 10 is TEM results for different salts.
Figure 11 is XRD results for different salts.
FIG. 12 is the results of electronic tongue testing of different salts.
Figure 13 is the results of the electronic nose test for different salts.
FIG. 14 is a morphological observation of pathological sections of liver tissues of various groups of mice.
Fig. 15 shows the inhibition of tyrosinase (monophenolase) in vitro by salt solutions of different concentrations, indicating that there is a very significant difference (p <0.01) between the experimental groups compared to the original salt, n 3.
Fig. 16 shows the inhibition of tyrosinase (diphenolase) in vitro by different salt solutions, indicating that there is a very significant difference (p <0.01) between the experimental groups compared to the original salt, and n is 3.
FIG. 17 is a graph of the effect of 0.5% mass fraction of each sample on B16 cell migration, a-Normal; b-a primary salt; c-arbutin; d-pine salt; e-bamboo salt; f-plum salt.
FIG. 18 is a graph of the effect of 0.5% mass fraction of each sample on the B16 cell cycle, a-Normal; b-a primary salt; c-arbutin; d-pine salt; e-bamboo salt; f-plum salt.
FIG. 19 is a graph showing the change in body weight of the experimental animals in FIG. 6.3.1.
FIG. 20 is the blood pressure change in the experimental rat of 6.3.2.
The present inventors have conducted extensive and intensive studies for a long time and have unexpectedly prepared a non-bamboo plant salt having excellent biological effects by subjecting a mixture of non-bamboo extracts (e.g., plum extract and/or pine extract, etc.) which have not been prebaked, and crude salt to a high temperature treatment at a temperature higher than the melting point of the crude salt (e.g., 800-. The preparation method of the non-bamboo plant salt is simple and is easy to popularize on a large scale. On this basis, the inventors have completed the present invention.
Term(s) for
In the present invention, the term "functional plant salt" refers to non-bamboo plant salt, and refers to a product prepared from non-bamboo extract as a plant raw material. Preferably, the non-bamboo plant salt is pine salt and/or plum salt.
In the present invention, the term "without pre-baking" means that the raw material for preparing the non-bamboo plant salt is a direct mixture of non-bamboo extract and raw salt, and the mixture is not subjected to one or more low-temperature (e.g., below 800 ℃) pre-baking treatments for traditionally preparing bamboo salt.
In the present invention, the term "plant extract" is a non-bamboo extract.
Non-bamboo extract
In the present invention, the term "non-bamboo extract" refers to an extract of a substance selected from the group consisting of: various fruits, vegetables, edible flowers, Chinese herbal medicines and medicinal and edible plants, preferably extracts of substances selected from the group consisting of: plum, pine, tea, lotus, chrysanthemum, pomegranate, crabapple, edible fungus, algae, and preferably plum extract and/or pine extract.
Said plum extract is selected from the group consisting of: plum extract, plum leaf extract, plum branch extract, plum root extract, plum kernel extract, or a combination thereof.
The pine extract is selected from the group consisting of: pine needle extract, pine bark extract, pine pollen extract, or a combination thereof.
Non-bamboo plant salt and preparation method and application thereof
The invention discloses a functional plant salt and a preparation method and application thereof, relates to the fields of food, seasonings, daily chemicals, medicines and the like, and aims to provide a natural, safe and nutritional plant salt with multiple biological health care effects. The method is characterized in that a medium-frequency smelting technology or a resistance heating technology is adopted, plant extracts of different types and different sources and edible salt are uniformly mixed according to a certain mass ratio (0-40%), then the mixture is placed into a medium-frequency smelting furnace or a resistance furnace, the temperature is increased to 800-1400 ℃ under normal pressure for melting, and crystallization is carried out after cooling, so that plant salt series with different colors, different physicochemical properties, different lattice structures, different flavors and different purposes are obtained. The method is characterized in that the salt solution is changed from weak acidity to strong basicity, the color is changed from white to different colors such as gray, light pink, purple red, bright blue and the like, and the salt solution presents crystal clear clean appearance and is accompanied with light sulfur taste; the unit cell size of the salt is smaller than that of the original salt, the element composition is richer, the potassium content is improved by about 100 times, the calcium content is improved by about 7 times, and other beneficial elements are also obviously increased. The health efficacy of functional plant salts include, but are not limited to: the whitening, liver protecting, stomach invigorating, detoxifying, anti-inflammatory, anti-allergic, oral cavity cleaning, hypertension preventing, migraine treating and intestinal canceration inhibiting effects, can be widely applied to food, beverage, seasoning, health food, medicine, daily chemicals, personal care products and the like, and has a very wide prospect. The melting point of the crystal lattice of the sodium chloride is 801 ℃, so that the temperature for completely melting the salt is at least about 900 ℃, and the traditional kiln can not meet the temperature requirement generally. Intermediate frequency smelting and resistance heating are two technologies capable of efficiently converting electric energy into heat energy, can quickly reach the smelting temperature of 900-1700 ℃, have high production efficiency, and can realize mechanized, automatic, large-scale, standardized and clean production. Meanwhile, functional plant salt series with different colors, stable performance, excellent quality and standardized quality can be obtained by controlling the variety, the adding proportion, the smelting temperature, the heat preservation time and the temperature rising and falling speed of the plant extract.
The invention creatively applies the medium-frequency melting technology or the resistance heating technology, mixes the salt with different plant extracts, and then melts the mixture at the high temperature of 800-1400 ℃, thereby forming the series of functional plant salts with different colors, different lattice structures, different physicochemical indexes, different flavors and different purposes.
Medium frequency smelting is a commonly used metal refining means, i.e. a power frequency 50Hz alternating current is converted into a medium frequency (300-1000 Hz), a three-phase power frequency alternating current is rectified to be a direct current, the direct current is converted into an adjustable medium frequency current, the adjustable medium frequency current is supplied to a medium frequency alternating current flowing through a capacitor and an induction coil, high-density magnetic lines are generated in the induction coil, and a high temperature is rapidly produced through a magnetic conductive container (such as a graphite silicon carbide crucible), so that a material (namely a mixture of salt and an extract) is heated to be molten. The intermediate frequency smelting technology can reach the required smelting temperature in a short time, and the production efficiency is greatly improved.
The resistance heating technology is to convert electric energy into heat energy through a resistor body, thereby rapidly heating materials. The current commercial resistance furnace in China mainly uses molybdenum rods, silicon rods or carbon rods as heating elements, and the highest furnace temperature can reach 1700 ℃. The resistance heating technology can control the heating time, temperature, heating rate and the like, thereby achieving the accurate control of process parameters and the standardization of products.
The plant extract is also called phytochemicals (i.e. plant secondary metabolites), and refers to different precision preparations and characteristic monomeric compounds obtained by extracting, separating and purifying various fruits, vegetables, flowers, Chinese herbal medicines, medicinal and edible plants and new resource foods by different technical means, such as extracts from plum, orchid, bamboo, chrysanthemum, pine, lotus, tea, pomegranate, begonia, edible fungi, algae and the like.
The algae extract can be algae (such as herba Zosterae Marinae, thallus laminariae, Cyrtymenia Sparsa, etc.), or algae cultured in fresh water or sea fresh water, such as blue algae, green algae, chlorella, etc.; the edible fungus extract comprises extracts of Lentinus Edodes, Agaricus campestris, needle Mushroom, caulis Bambusae in Taeniam, Tricholoma, Auricularia, Tremella, Hericium Erinaceus, Ganoderma, truffle, and Boletus and its processing residue (such as Lentinus Edodes stem); the extracts derived from microorganisms include extracts of various yeasts (also called yeast extract), such as beer yeast, baker's yeast, etc.
The invention is mainly technically characterized in that: by adopting a medium-frequency smelting technology or a resistance heating technology, plant extracts from different sources and salt are uniformly mixed according to the proportion of 0-40% (extract powder is uniformly adsorbed on the surface of salt particles), the mixture is heated to 800-1400 ℃ under normal pressure for smelting together, and the mixture is cooled and recrystallized to obtain plant functional salt series with different colors, different lattice structures, different physicochemical indexes, different flavors and different purposes. According to the source of the plant extract, the difference of the effective components, the adding proportion, the smelting parameters and the like, the obtained novel plant salt series has multiple functions of whitening, protecting liver, invigorating stomach, detoxifying, resisting inflammation, resisting allergy, cleaning oral cavity, preventing hypertension, treating migraine, inhibiting intestinal canceration and the like, and can be widely applied to the fields of food, beverage, seasonings, health-care food, medicines, daily chemicals, personal care products and the like.
The invention has the following general processes and process parameters:
uniformly mixing the plant extract powder and the salt according to the mass ratio of 0-40% (preferably 5-25%);
putting the mixed material into a magnetic crucible and placing the magnetic crucible in an intermediate frequency smelting furnace, and starting to heat up at the heating rate of 1-200 ℃/min; or putting the mixed materials into a corundum crucible and placing the corundum crucible in a resistance furnace, and starting to heat up at the heating rate of 1-20 ℃/min;
heating to a certain temperature of 800-1400 ℃ under normal pressure, and keeping the temperature for 0-6 h;
pouring out the materials, and naturally cooling or controllably cooling;
cooling, crystallizing, crushing and sieving to obtain the product;
the heating temperature can be set in 1 or more intervals between 800 and 1400 ℃ according to the requirement;
the above mixing-melting-recrystallization-cooling-pulverization process may be carried out once or more times. (1) The raw material salt (also called raw salt) can be sea salt, lake salt, well salt and mineral salt, and preferably crude sea salt (namely solar salt). (2) According to the different parameters of the plant extract source, the adding proportion, the smelting temperature and the like, the health effect of the functional plant salt provided by the invention can be embodied as that the health effects of the non-bamboo functional plant salt (such as pine salt and plum salt) include but are not limited to: whitening skin, protecting liver, and preventing hypertension. The application fields of the product comprise food, beverage, condiment, health food, medicine, daily chemical, personal care product and the like. (3) In addition, the medium-frequency melting technology and the resistance heating technology can also be directly used for processing the salt, namely, no plant extract or other foreign substances are added in the manufacturing process, and the original salt is converted into the baked salt which has higher edible safety and more crystal-clear appearance.
The method has the outstanding advantages that the method adopts a medium-frequency smelting technology or a resistance heating technology, can mix the raw salt with the plant extracts (powder) of any source and in any proportion, recrystallizes the mixture after the mixture is melted in a precisely set temperature interval, obtains the salt product with special biological efficacy through one or more times of baking, and can realize large-scale industrial production with mechanization, automation, standardization and cleanness. The invention creates non-bamboo series plant functional salt represented by pine salt and plum salt, which is a revolution of salt industry (especially middle and high-end salt market) and has very important significance for the health industry of all mankind.
In order to solve the technical problems, the invention provides a preparation method of functional plant salt, which comprises the following steps:
one or more plant extract powders are evenly mixed with fine salt according to the addition amount of 1-40 percent and baked at the high temperature of 800-1400 ℃ for one time or several times to obtain the functional plant salt.
As an improvement of the preparation method of the functional plant salt of the invention:
the plant extract powder is one of the following components:
uniformly mixing the plant extract powder with fine salt according to the addition amount of 1-40%, and roasting at the high temperature of 800-1400 ℃ for one time to obtain the functional plant salt.
As a further improvement of the preparation method of the functional plant salt of the invention:
when the plant extract powder is plural (at least two types):
uniformly mixing various plant extract powders to obtain mixed powder;
the obtained mixed powder is evenly mixed with fine salt according to the addition amount of 1-40 percent and is roasted at the high temperature of 800-1400 ℃ for one time to obtain the functional plant salt.
As a further improvement of the preparation method of the functional plant salt of the invention:
when the plant extract powder is plural (at least two types):
s1, uniformly mixing the plant extract powder with fine salt according to the addition of 1-40%, roasting at the high temperature of 800-1400 ℃ and preserving heat for a period of time;
s2, taking out the product obtained in the step S1 at the temperature of 800-1400 ℃ for a certain time, adding another plant extract powder into the product after crushing, and repeating the steps S1 and S2 (note: baking is continued at the original temperature or higher) until all the plant extract powder is baked to obtain the functional plant salt.
In order to solve the technical problems, the invention also provides the functional plant salt prepared by the preparation method.
In order to solve the technical problems, the invention also provides the application of the functional plant salt:
the functional plant salt is used for pharmaceutical aids, health foods/drinks, functional foods/drinks, table seasoning salt, cooking salt, special-purpose cosmetics or personal care products.
The functional plant salt is characterized in that plant extracts from different sources and salt are mixed uniformly according to a certain proportion by adopting a medium-frequency smelting technology or a resistance heating (also called ohmic heating) technology, then the mixture is smelted at the temperature of 800-1400 ℃ under normal pressure, and the cooled mixture is recrystallized to obtain novel health salt, which can be widely applied to foods, drinks, condiments, health-care foods, medicines, daily chemicals, personal care products and the like. The non-bamboo plant functional salt which is originally created by the invention and represented by pine salt and plum salt has the same biological effect as bamboo salt, and also has various effects of preventing hypertension, whitening skin, protecting liver and the like.
According to the difference of the species, the source and the effective components thereof, the adding proportion, the baking process parameters and the like of the plant extract, the plant functional salt with different colors, different lattice structures, different physicochemical indexes, different flavors and different purposes can be obtained and can be divided into two series of bamboo salt and non-bamboo salt. The outstanding characteristics and commonality of the product are as follows: the salt solution is changed from weak acidity to strong basicity, the color is changed from white to different colors such as grey, light pink, purple red, bright blue and the like, and the salt solution presents crystal clear clean appearance and is accompanied with light sulfur taste; the unit cell size of the salt is smaller than that of the original salt, the element composition of the salt is richer, the potassium content is improved by about 100 times, the calcium content is improved by about 7 times, and other beneficial elements are also obviously increased.
The medium-frequency smelting technology is characterized in that a power frequency 50Hz alternating current is converted into a medium frequency (300-1000 Hz), a three-phase power frequency alternating current is rectified to be converted into a direct current, the direct current is converted into an adjustable medium frequency current, the adjustable medium frequency current is supplied to a medium frequency alternating current flowing through a capacitor and an induction coil, high-density magnetic lines are generated in the induction coil, and a magnetically conductive container (such as a graphite silicon carbide crucible) is used for rapidly producing high temperature so as to heat a material (namely a mixture of salt and an extract) to melt the material. The smelting temperature can be quickly reached in a short time, and the production efficiency is greatly improved.
The specific process and parameters are as follows:
uniformly mixing different plant extract powder and salt according to the mass ratio of 0-40% (preferably 5-25%);
putting the mixed material into a magnetic crucible, putting the crucible into a medium-frequency smelting furnace, and starting to heat at a heating rate of 20-100 ℃/min;
raising the temperature to a certain temperature range between 800 and 1400 ℃ under normal pressure, and preserving the heat for 0 to 6 hours;
pouring out the molten material, and naturally cooling or controllably cooling;
cooling, crystallizing, crushing and sieving to obtain the product;
the smelting temperature can be set in 1 or more intervals between 800 and 1400 ℃;
the above mixing-melting-cooling-recrystallizing-pulverizing process can be carried out once or more times.
The resistance heating (also called ohmic heating) technology is to convert electric energy into heat energy so as to heat the material quickly. Compared with a flame furnace, the resistance furnace has high thermal efficiency and easy temperature control, and is divided into a direct heating mode and an indirect heating mode. At present, most resistance furnaces are heated indirectly, and the devices are equipped with resistance elements for realizing the electric-thermal conversion, called electrothermal bodies, which transfer the heat energy to the materials to be processed. The commonly used electric heating body comprises a silicon carbide rod and a molybdenum disilicide rod, wherein the temperature of the silicon carbide rod can reach 1400 ℃, and the temperature of the silicon molybdenum rod can reach 1700 ℃.
The specific process and parameters are as follows:
uniformly mixing different plant extract powder and salt according to the mass ratio of 0-40% (preferably 5-25%);
putting the mixed material into a corundum crucible, putting the corundum crucible into a resistance furnace, and starting to heat up at the heating rate of 1-20 ℃/min;
heating to a certain temperature of 800-1400 ℃ under normal pressure, and keeping the temperature for 0-6 h;
pouring out the materials, and naturally cooling or controllably cooling;
cooling, crystallizing, crushing and sieving to obtain the product;
the smelting temperature can be set in 1 or more intervals between 800 and 1400 ℃;
the above mixing-melting-recrystallization-cooling-pulverization process may be carried out once or more times.
The plant extract is also called phytochemicals, namely a secondary metabolite of the plant. Comprises various fruits, vegetables, flowers, Chinese herbal medicines, medicinal and edible plants, plant crude extracts of new resource foods derived from plants, high-precision preparations and monomer compounds, such as bamboo, plum, pine, tea, lotus, chrysanthemum, pomegranate, Chinese flowering apple, edible fungi, algae extracts and the like. The same plant source can also be extracts of different parts, such as bamboo extract including folium Bambusae extract (such as bamboo leaf flavone and bamboo leaf polyphenol), caulis Bambusae in Taenia extract (such as bamboo shavings triterpene and bamboo shavings polysaccharide), caulis Bambusae in Taenia extract, bamboo shoot extract (such as bamboo shoot sterol and bamboo shoot amino acid peptide), mume fructus extract including mume fructus extract, mume fructus leaf extract, mume fructus branch extract, mume fructus root extract and mume fructus kernel extract, and pine extract including folium Pini extract, cortex Pini extract and pollen Pini extract.
According to the variety and source of plant extracts, plant functional salt is divided into two series of bamboo and non-bamboo. The functional salt is classified as bamboo functional salt prepared from bamboo extract series, and non-bamboo functional salt prepared from extract (or called extract) of algae, mushroom (edible fungi) and microorganism.
The salt raw material (i.e. raw salt) can be sea salt, lake salt, well salt and mineral salt, and preferably is crude sea salt (also called solarized sea salt).
The health efficacy of the non-bamboo functional salt includes but is not limited to: preventing hypertension, whitening skin, and protecting liver.
The application fields of the functional plant salt comprise: food, beverage, seasoning, health food, medicine, daily chemical and personal care product.
The intermediate frequency melting technology or the resistance heating technology can also be used for directly processing the salt (namely, no plant extract or any other foreign substances are added), and the salt is converted into the roasted salt with higher edible safety.
Compared with the prior art, the invention has the following technical advantages:
the specific biological functions of the functional plant salt prepared by the invention comprise: bacteriostasis, anti-inflammation and antivirus; protecting liver, invigorating kidney, tranquilizing mind, invigorating stomach, removing toxic substances, relieving alcoholic intoxication, stopping bleeding, and eliminating hemorrhoid; treating migraine, allergic rhinitis, anti-inflammatory effect on human gingival fibroblast, reducing weight, protecting oral health, inhibiting colon canceration, increasing anti-mutation and in vitro anticancer effects, resisting mutation activity and in vitro anticancer effect of HepG2 human liver cancer cell, and preventing nerve cell apoptosis. In addition, the skin care product is beneficial to improving the permeability of epidermal cells of a human body and helping skin care factors in daily chemicals to better play a role.
The specific application of the functional plant salt prepared by the invention comprises the following steps: a pharmaceutical adjuvant; health food/drink; functional food/beverage products; table seasoning salt, cooking salt; cosmetics for special use, personal care products (skin cream, foundation, soap, pack, toothpaste, mouthwash, shampoo, bath salt, etc.); the final product can be in the form of powder, capsule, tablet, granule, effervescent tablet, water solution, emulsion, spray, etc.
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are by weight.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.
Universal feedstock and apparatus
Pine needle extract: brown powder, available from qingya biotechnology limited of shanxi;
bamboo leaf extract (bamboo leaf flavone): brown powder, available from santa se, zhejiang, biotechnology limited (angji);
plum branch extract: yellow powder, provided by Youmett Biotechnology, Inc. of Hangzhou;
raw salt (sun sea salt): from Shandong Weifang, the actually measured NaCl content is 93.36%;
intermediate frequency smelting furnace: the heating element is a red copper induction coil, and the crucible is a magnetic graphite silicon carbide crucible;
a tubular resistance furnace: the heating element is a silicon carbide rod.
Example 1, a method for preparing functional plant salt, comprising the steps of:
1. obtaining a plant extract powder:
the plant extracts include, but are not limited to, plant extracts of various fruits, vegetables, edible flowers, Chinese herbal medicines and medicinal and edible plants, and new resource foods derived from plants, such as bamboo, plum, pine, tea, lotus, chrysanthemum, pomegranate, crabapple, edible fungi, algae extracts, and the like.
The same plant source can also be extracts of different parts, such as bamboo extract including folium Bambusae extract (bamboo leaf flavone, bamboo leaf polyphenol), caulis Bambusae in Taenia extract, caulis Bambusae in Taeniam extract, and bamboo shoot extract, mume fructus extract including mume fructus extract, mume fructus branch extract, and mume fructus root extract, and pine extract including folium Pini extract, cortex Pini extract, and pollen Pini extract.
2. Uniformly mixing the plant extract powder in the step 1 with fine salt according to the addition of 1-40% to obtain a mixture;
putting the mixture into a special container, heating to 800-1400 ℃ in a medium-frequency smelting furnace (or other mechanized ovens) at a certain heating rate to obtain the functional plant salt.
The fine salt can be sea salt, lake salt, well salt, and mineral salt, preferably crude sea salt.
The above percentages are mass ratios.
Example 2, the plant extract powder in example 1 is changed into at least two plant extract powders, that is, after at least two plant extract powders are uniformly mixed according to any proportion, mixed powder is obtained; uniformly mixing the mixed powder with fine salt (sieved) according to the addition amount of 1-40% to obtain a mixture; the rest is equivalent to embodiment 1.
Example 3, the plant extract powder of example 1 was changed to at least two plant extract powders, the specific steps were as follows:
1. mixing a plant extract powder with fine salt to obtain a mixture;
2. putting the obtained mixture into a special container, heating to 800-1400 ℃ in a medium-frequency smelting furnace (or other mechanized ovens) at a certain heating rate, and preserving heat for a certain time to obtain an obtained substance;
3. taking out the product obtained in step 2 and keeping at 800-1400 deg.C for a certain time, pulverizing, adding another plant extract powder, and repeating steps 1-3 (note: baking at original temperature or higher) until all plant extract powder is baked to obtain functional plant salt
Note: the total addition amount of the plant extract powder is 1-40% (mass ratio).
In conclusion, the functional plant salt can be baked at one time after the plant extract is added, or can be baked by adding and grading.
In the high-temperature smelting process, due to the infiltration of different mineral elements in different plant extracts, the element composition, the NaCl crystal lattice and the crystal configuration are obviously changed. According to different adding proportions and baking modes, the food takes on different colors, such as gray, purple, pink, white, blue, turquoise and the like; the pH value of the 5% saline solution is increased from 6-7 of the raw material salt to 8-12; the total amount of NaCl is reduced, and the content of other elements (especially various trace elements) is increased; the taste is changed from pure salty taste to rich and mellow salty, fresh and sweet taste, and the taste is still slightly smelly preserved eggs.
For example, "pine salt", "bamboo salt" and "plum salt" obtained by adding pine, bamboo and plum extracts have significant differences in physicochemical properties, appearance, color, flavor and taste of the obtained plant salt due to differences in sources, chemical compositions and addition amounts of the extracts, and differences in baking times and process parameters, and thus have different biological effects.
Specific biological functions of functional plant salts include: bacteriostasis, anti-inflammation and antivirus; protecting liver, invigorating kidney, tranquilizing mind, invigorating stomach, removing toxic substances, relieving alcoholic intoxication, stopping bleeding, and eliminating hemorrhoid; treating migraine, allergic rhinitis, anti-inflammatory effect on human gingival fibroblast, reducing weight, protecting oral health, inhibiting colon canceration, increasing anti-mutation and in vitro anticancer effects, resisting mutation activity and in vitro anticancer effect of HepG2 human liver cancer cell, and preventing nerve cell apoptosis. In addition, the skin care product is beneficial to improving the permeability of epidermal cells of a human body and helping skin care factors in daily chemicals to better play a role.
Specific uses of functional plant salts include: as a pharmaceutical adjuvant; health food/drink; functional food/beverage products; table seasoning salt, cooking salt; cosmetics for special uses, and personal care products (skin cream, foundation lotion, soap, face pack, toothpaste, mouth wash, shampoo, bath salt, etc.); the final product can be in the form of powder, capsule, tablet, granule, effervescent tablet, water solution, emulsion, spray, etc.
In summary, the invention provides a preparation method of functional plant salt, which is characterized in that different plant extracts and salt are uniformly mixed according to a certain proportion and then baked once or for several times at the high temperature of 800-1400 ℃ to form the functional plant salt with different colors, different spatial structures, different physicochemical properties, different flavors and different purposes. The functional plant salt prepared by the invention has a health-care effect, is beneficial to improving the permeability of epidermal cells of a human body and helping skin-care factors in daily chemicals to better act, and can be used as a pharmaceutical aid, a health-care food/beverage, a functional food/beverage, table seasoning salt, cooking salt, a special-purpose cosmetic or a nursing product.
Example 4 preparation of vegetable salt-pine salt (intermediate frequency melting)
Weighing 6.0g of pine needle extract and 24.0g of raw Salt respectively, uniformly mixing, placing into a magnetic conduction crucible, placing into a medium frequency smelting furnace, heating to 1000 ℃ at normal pressure at a heating rate of 100 ℃/min, keeping the temperature at 1000 ℃ for 1h, taking out, and naturally cooling to obtain pink-purple pine Salt (the code is Plum Salt), wherein the color and appearance of the pine Salt are shown in figure 1.
Example 5 preparation of vegetable functional salt, New purple bamboo salt (intermediate frequency melting)
Respectively weighing bamboo leaf extract 10g and original salt 40g, mixing well, placing into magnetic crucible, placing into intermediate frequency smelting furnace, heating to 900 deg.C at normal pressure at a heating rate of 100 deg.C/min, keeping the temperature for 1.5h, taking out, naturally cooling, and pulverizing to obtain new purple bamboo salt with color and appearance shown in FIG. 2.
Example 6 preparation of vegetable functional salt plum salt (intermediate frequency melting)
Respectively weighing 5.0g of Plum branch extract and 25.0g of raw Salt, uniformly mixing, putting into a magnetic crucible, putting into a medium-frequency smelting furnace, heating to 1300 ℃ at normal pressure at a heating rate of 100 ℃/min, keeping the temperature at 1300 ℃ for 1h, taking out, and naturally cooling to obtain blue Plum Salt (marked as Plum Salt), wherein the color and appearance of the Plum Salt are shown in figure 3.
Example 7 preparation of vegetable salt-pine salt (resistance heating)
Weighing 4.0g of pine needle extract and 26.0g of raw salt respectively, mixing uniformly, placing into a magnetic crucible, placing into a tubular resistance furnace, heating to 1100 ℃ at normal pressure at a heating rate of 100 ℃/min, keeping the temperature at 1100 ℃ for 2h, taking out, and naturally cooling to obtain pink-purple pine salt, wherein the color and appearance of the pine salt are shown in figure 4.
Example 8 preparation of vegetable functional salt, New purple bamboo salt (resistance heating)
Respectively weighing 15g of bamboo leaf extract and 35g of original salt, mixing well, placing into a corundum crucible, placing into a tube furnace, heating to 950 ℃ at normal pressure at a heating rate of 10 ℃/min, keeping the temperature for 2.5h, taking out, naturally cooling, and pulverizing to obtain the purple bamboo salt, wherein the color and appearance of the purple bamboo salt are shown in figure 5.
Example 9 preparation of vegetable functional salt plum salt (resistance heating)
Respectively weighing 8.0g of plum branch extract and 22.0g of raw salt, mixing well, placing into a corundum crucible, placing into a tubular resistance furnace, heating to 1250 ℃ at normal pressure at a heating rate of 10 ℃/min, keeping the temperature at 1250 ℃ for 1.5h, taking out, and naturally cooling to obtain blue plum salt, wherein the color and appearance of the plum salt are shown in figure 6.
Performance test (in the experiment, pine, bamboo and plum salt are respectively samples obtained in examples 4, 5 and 6)
1.1 color difference analysis
The type, the adding proportion, the smelting temperature, the heat preservation time and the temperature rising rate of the plant extract can directly influence the color and the appearance of the final product. FIG. 7 shows that the pine salt obtained in example 4, the bamboo salt obtained in example 5, and the plum salt obtained in example 6 can respectively show different colors such as gray, light pink, bluish violet, and bright blue by adding 5 to 35% of the original salt of different plant extracts and melting at 800 to 1400 ℃ for 1 to 2 hours under normal pressure. Meanwhile, as can be seen from fig. 7, the roasted salt obtained by smelting the raw salt without any foreign substances at 800-900 ℃ for 1-2 hours has higher transparency and brighter color.
The chromatometer is a common photoelectric integral type color measuring instrument, it adopts light receiver capable of sensing red, green and blue colors, utilizes the standard light source in the interior of instrument to irradiate measured object, and makes amplification treatment of the light current sensed by each light receiver, and makes once integral measurement in the whole visible light wavelength range to obtain tristimulus value and chromaticity coordinate of the transmitted or reflected object color so as to obtain the signal of said color, and utilizes the system to give out the color difference value between the measured samples.
The main indexes measured by the color difference meter comprise L, a and b. Wherein,
l denotes the brightness of the sample, with a larger value indicating a brighter;
a represents the red and green directions, "+" represents a bias towards red, and "-" represents a bias towards green;
b represents a blue-yellow phase, "+" indicates a shift towards yellow, and "-" indicates a shift towards blue.
The Δ L value represents the difference between the sample value and the reference point; the delta a value represents the difference of a value of the sample relative to a reference point, and can better represent the red-green value of the sample.
In the present invention, the reference point refers to a value of a standard sample, and the present invention uses a standard white board as the standard sample.
Table 1 is a summary of the color difference analysis data for different plant functional salts. It can be seen that the Δ L values of the raw salt and the roasted salt are 84.18 + -0.25 and 82.83 + -3.47, respectively, showing that they are nearly white; and the delta L value of other plant functional salts is between 25 and 50, wherein the darkest is the mangosteen salt, and the brightest is the plum salt. The a and b values of different samples differed significantly (p <0.05), indicating that the color directions differed greatly and distinct hues were exhibited.
TABLE 1 color value parameters of different plant functional salt samples
1.2 change in pH
The pH value of food has an important influence on the maintenance of acid-base balance in the human body, and is therefore classified into acidic food and alkaline food. In modern society, the dietary structure of the public is generally acidic, and acidic constitutions can be formed over time, so that the immunity is reduced and the sub-health state is caused. The pH values of different salt samples at a concentration (w/v) of 0.2-5% were measured, as shown in Table 2.
The original salt solution with the concentration of 1 percent or below is weakly acidic (the pH value is less than 7), and the original salt solution is obviously increased after being baked; whereas all plant functional salt solutions exhibit strong alkalinity (pH >9) even at 0.2% concentration, the difference is quite significant compared to the original salt (p < 0.001). It was shown that a large amount of alkali metal elements were incorporated into the lattice structure of NaCl or attached to the surface of the original salt after it was co-melted with the plant extract.
TABLE 2 pH values of different plant functional salt samples
1.3 Change in conductivity
Conductivity data were obtained for different samples at 0.2-5% concentration (w/v) as shown in Table 3. As can be seen from the table, the conductivity of the plant functional salt of the present invention is not significantly different from that of the original salt at a salt solution concentration (< 1.0%) which is tolerable to human body.
TABLE 3 conductivity values of different plant functional salt samples
1.4 atomic Spectroscopy
The atomic energy spectrum not only can scan and shoot the surface morphology of the material, but also can detect various elements on the surface of the material and the relative contents of the elements. The plant functional salt and the control sample (particle) thereof prepared by the invention are placed under an atomic energy spectrometer (SU-8010, Japan Hitachi Co., Ltd.) to carry out qualitative, semi-quantitative and quantitative analysis (detection limit is 0.5%) on the micro-area points, lines and surfaces of the surface layer of the sample, and the obtained results are shown in Table 4.
TABLE 4 elemental composition (%)
1.5 elemental composition analysis
The element detection was entrusted to the Western-An national Union quality detection technology, Inc., and the results are shown in Table 5.
The main component of the salt is sodium chloride, and in addition, the salt also contains a small amount of other mineral elements. The salt with higher refining degree has higher sodium chloride content and less other nutrient elements, which is unfavorable for human health.
The potassium element can regulate the intracellular osmotic pressure and the acid-base balance of body fluid, participate in the metabolism of intracellular sugar and protein, contribute to maintaining the nerve health and normal heartbeat rule, prevent stroke and assist the normal contraction of muscles. In the case of hypertension caused by ingestion of high sodium salt, potassium salt or potassium-rich food has a hypotensive effect.
Magnesium is the main cation in human cells, and can affect the transport of potassium ions and calcium ions, regulate and control the transmission of signals, participate in energy metabolism, the synthesis of proteins and nucleic acids, activate and inhibit catalytic enzymes and the like.
Phosphorus is one of the basic components constituting nucleic acid of genetic materials, and phosphorus and calcium are important constituent materials of bones and teeth. Phosphorus can keep balance of ATP metabolism in vivo, regulate acid-base balance in vivo, and participate in metabolism of energy in vivo.
Manganese is a coenzyme for SOD, and its deficiency can cause neurasthenia, affect intellectual development, and also cause a decrease in insulin synthesis and secretion, affecting carbohydrate metabolism.
Molybdenum is an essential trace element for human body, is one of the basic components of xanthine oxidase and aldehyde oxidase in liver and intestine of animal body, and is also a component of heparin sulfite oxidase. Research shows that molybdenum has obvious caries preventing effect and strong urinary calculus inhibiting effect, and may be used to treat kidney calculus caused by molybdenum deficiency.
Vanadium is a trace element essential to the human body and is believed to contribute to the prevention of cholesterol accumulation, the reduction of blood glucose, the reduction of blood pressure, the prevention of dental caries, the help of the production of red blood cells, and the like.
Selenium is an important trace element necessary for human bodies, the Chinese nutrition society ranks selenium as one of 15 nutrients necessary for human bodies, generally, the selenium is considered to be closely related to immunity, aging, reproductive function and the like, and the selenium deficiency is an important reason of keshan disease.
Chromium is also an essential micronutrient, plays an important role in all insulin-regulating activities, and is an important blood glucose regulator. Meanwhile, chromium deficiency is also associated with the formation of myopia.
Silicon is mainly concentrated in bones, lungs, lymph nodes, pancreas, adrenal glands, nails and hair, has the highest content in connective tissues such as aorta, trachea, tendons, bones, skin and the like, and has special significance for the bone health of human bodies. However, due to the development of industry and the change of life style, the silicon element in modern human body is often extremely insufficient.
From the element detection results of different salt samples, the plant functional salt and the original salt (and the roasted salt thereof) have quite obvious difference. The potassium content of pine, bamboo and plum salt is remarkably increased, especially the potassium element in the novel bamboo salt obtained in the embodiments 2 and 3 is more than 100 times of that of the original salt, and is 14.9 percent higher than that of the benevolence mangosteen salt produced by the traditional process; the bamboo salt (including 2 samples of new and old bamboo) contains magnesium, iron, nickel, zinc, molybdenum and vanadium which are obviously higher than those of original salt and other plant salt, and the pine salt and the plum salt obtained by the new technology also have the selenium. In all samples, the plum salt possessed much higher contents of iodine (28mg/kg), boron (26.6mg/kg), phosphorus (424mg/kg), chromium (4mg/kg) and manganese (18mg/kg) next to bamboo salt than the other samples. The silicon content of the plant salt is obviously higher than that of the original salt and the roasted salt, and the bamboo salt produced by the new process is higher than that of the traditional produced Renshan bamboo salt. Meanwhile, selenium element is also detected in the pine salt and the plum salt.
The bamboo salt also contains a certain amount of sulfide, and has typical odor of preserved egg. Hydrogen sulfide has various neuroprotective effects, which are proved by a large number of experiments at home and abroad, and mainly comprise anti-neuritis reaction, anti-oxidative stress damage and anti-hypoxia and anti-ischemic nerve damage, and can increase the plasticity of nerve cells, thereby reducing the apoptosis of hippocampal neurons.
In addition, particularly, the edible safety of the salt is greatly improved by high-temperature smelting at the temperature of more than 800 ℃, and the content of heavy metals (lead, arsenic and mercury) in the roasted salt is greatly reduced compared with that in the original salt. Meanwhile, other harmful components (such as organic plastic particles and the like) in the original salt are also completely removed.
TABLE 5 results of elemental detection analysis of different samples
Remarking: the detection limit of the phosphorus element is 20 mg/kg; the detection limit of molybdenum, antimony, selenium, tin and total arsenic elements is 0.01 mg/kg; the detection limit of cadmium and germanium elements is 0.001 mg/kg; the detection limit of thallium element is 0.0001 mg/kg.
2.1 micrographs of the appearance of the samples
The appearance of the sample obtained from the plant functional salt particles obtained in the practice was photographed under the condition of 10 times of the eyepiece and 5 times of the objective lens by a stereo microscope (Jiangnan brand SE220) and is shown in FIG. 8.
It can be seen from FIG. 8 that the original salt and roasted salt are substantially white, the particle surface is smooth, and the appearance is regular. While pine salt, bamboo salt and plum salt are different in color, have larger crystal arrangement difference and are more glossy. The color of the new bamboo salt prepared by the invention is similar to that of the mangosteen salt, but the new bamboo salt is purer and has glittering and translucent appearance, and the new bamboo salt has more impurities.
2.2 observation results of scanning Electron microscope
The plant functional salts of the examples were morphologically characterized by means of a GeminiSEM 300 scanning electron microscope from the company GmbH of Card Zeiss Microcopy, Germany. The different sample particles were observed under an electron microscope, respectively, to obtain FIG. 9. The surface of the original salt and the baked salt is smooth and has no more attachments. The surface of the plant functional salt prepared by the invention is in the shape of sweet potato, and a plurality of small particles are attached to the surface of the particles.
2.3 observation results of Transmission Electron microscopy
The different salt samples were suspended in absolute ethanol (5%, w/v) and the plant functional salt particles obtained in the examples were morphologically characterized using a Transmission Electron Microscope (TEM), JEOL, JEM-1230, Japan. The transmission electron microscope images of the plant functional salts represented by pine salt, bamboo and plum prepared in examples 4, 5 and 6 are shown in fig. 10, the original salt is regular in shape and has a cubic regular structure of sodium chloride, while the plant functional salts have no specific rules and are attached with a few tiny particles around the original salt.
2.4 results of X-ray diffraction analysis
Pine salt obtained in example 4, bamboo salt obtained in example 5, plum salt obtained in example 6 and a control sample thereof were ground into powder (100-mesh sieve) and analyzed by an X-ray diffractometer (Bruker D8Advance) manufactured by Bruker, Germany. Because the crystal is composed of unit cells formed by regularly arranged atoms, X-rays scattered by different atoms interfere with each other to generate strong X-ray diffraction in certain special directions, the orientation and the intensity of the spatial distribution of the diffraction lines are closely related to the crystal structure, and the diffraction pattern generated by each crystal reflects the atom distribution rule in the crystal. The phase analysis, the unit cell parameter determination and the diffraction line intensity analysis can be carried out on the sample.
2.2.1 Crystal Structure analysis
Phase analysis is the most used aspect of X-ray diffraction among metals and is divided into qualitative analysis and quantitative analysis. The former compares the lattice spacing and diffraction intensity measured for the material with the diffraction data of a standard phase to determine the phase present in the material. The precise cell parameter data can reflect subtle differences in structure between different samples of a substance, or subtle changes in the structure of a crystal under the influence of external physicochemical factors. The unit cell parameters of several phases of the plant functional salts obtained in the examples are as follows:
TABLE 6 sodium chloride content and lattice parameter in different samples
TABLE 7 Potassium chloride content and lattice parameter in different samples
TABLE 8 Crystal size of salts
2.2.2X-ray diffraction spectra and simulated crystal structure diagrams
The unit cell patterns of the different salt samples were modeled by unit cell parameters. It can be seen from fig. 11 that the original salt is regular and cubic, while the pine salt has sharp ends and round bamboo salt, and the plum salt has an octahedral cone, which indicates that the crystal structure of the original salt and different plant extracts is greatly changed after high-temperature melting.
The electronic tongue is used for simulating the human tongue to analyze, recognize and judge a sample to be detected, and the obtained data is processed by a multivariate statistical method, so that the overall quality information of the sample is quickly reflected, and the recognition and classification of the sample are realized. The method is a detection technology which is based on a multi-sensor array, senses the overall characteristic response signal of a sample, and performs analog identification and quantitative and qualitative analysis on the sample. Mainly comprises a taste sensor array, a signal acquisition system and a mode recognition system 3.
The electronic nose is also called as a smell scanner, and is a novel instrument for quickly detecting food developed in the 90 s of the 20 th century. It uses special sensor and pattern recognition system to provide the whole information of tested sample quickly, indicating the implicit characteristics of sample. It is an instrument consisting of a selective electrochemical sensor array and a suitable identification method, which can identify simple and complex odours and can obtain results consistent with human sensory evaluations.
The Discrimination Index (DI) is a measure for determining the overall Discrimination effect of the sample by the electronic tongue and the electronic nose, and the range of the DI is [ -100, +100 ]. 100,0 means that it cannot distinguish taste from smell of the sample; +100 means that it can be distinguished effectively, and the closer the value is to 100, the better the distinguishing effect is.
3.1 taste traits Using electronic tongue
The plant functional salts obtained in the examples were prepared into 10% solutions, and their taste difference was examined by using electronic tongue (Smartongue, Isensogroup Co.).
From FIG. 12, it can be seen that the DI value of the electronic tongue reached 99.57, indicating that there is a significant difference in taste between different plant functional salts, which can be clearly distinguished. The contribution rate of the principal component 1(PC1) is 78.26%, which is much greater than that of the principal component 2(PC 2); the farther the distance on the abscissa is, the greater the difference is represented; the closer distance of the abscissa of the pine salt obtained in example 4/the bamboo salt obtained in example 5 and the salt of Arundina nucifera indicates that the two are closer in taste and both are significantly different from the original salt and the plum salt obtained in example 6.
3.2 olfactory characteristics by electronic nose
The electronic nose collects odor signals of different samples through 14 sensors, and then the data are processed by software to carry out principal component analysis, the plant functional salt obtained in the example is prepared into 10% solution, and the taste difference of the plant functional salt detected by the electronic nose (iNose, Isensogroup company) is shown in figure 13: the two main components which account for the most total information in the sample are respectively used as an abscissa PC1 and an ordinate PC2, the contribution rate of the main component 1(PC1) is 99.80 percent and is far greater than that of the main component 2, therefore, the farther the distance of the data of the two samples on the abscissa is, the larger the difference is, the abscissas of the plant functional salts in the graph are very close, and the three plant salts obtained by the method are relatively close to each other in olfaction. The smell of the bamboo salt (Renshan bamboo salt) prepared by the traditional method is closer to that of the original salt. In addition, according to the results of atomic energy spectrum analysis in table 4, sulfur elements are found on the surface of the bamboo salt, so that it can be explained that the bamboo salt has a light hydrogen sulfide smell, and the sulfur elements of other salts all reach the detection limit.
4.1 animal grouping and Experimental design
4.1.1 Experimental animals and groups
45 healthy male mice, SPF grade C57BL/6, were 5 weeks old, were acclimatized for one week and then randomized into 9 groups, with experimental design and dose configuration as shown in table 9. Except for normal control group (1)#) Except for the conventional feed, the rest 8 groups all use the salt-free basic feed. During the administration period, the oral dosage of the mice is obtained according to the recommended daily salt intake of 5g per person of the world health organization and a standard animal equivalent dose reduction algorithm, except 1#In addition, each gavage sample contained 4% NaCl and was gavage 1 time daily for 21 days. The temperature of the breeding environment is (22 +/-2) DEG C, and the relative humidity is 50-70%.
TABLE 9 animal test groups and dosages administered
To ensure comparability of the test data, normal control group 2#And model group 3#The dietary salts are all pure NaCl
4.1.2 acute hepatic injury modeling
Model control group and test group 1h after the last administration (3)#~9 #) Mice were all intraperitoneally injected with 0.1% CCl4Olive oil solution, CCl4BW at a dose of 10mL/kg, normal control group (1)#~2 #) An equal volume of olive oil was given and fasted overnight.
4.1.3 sample Collection and Biochemical index measurement
In the case of CCl injection4After 16h of olive oil solution, the mice were sacrificed by dislocation of the spine, the eyeballs were immediately removed to draw blood, and the mice were dissected to take the liver.
Centrifuging the blood sample at 3500r/min for 10min, and sucking the upper serum for blood biochemical index determination. After the whole liver was removed, 1 part of 0.4g of liver tissue was harvested, and put into a freezing tube containing PBS (pH 7.4) to homogenize, and the remaining liver tissue was put into a freezing tube and stored in a refrigerator at-80 ℃ for use. Homogenizing 0.4g of the sheared liver, centrifuging at 4000r/min for 20min in a ultra-low temperature centrifuge, collecting the supernatant, and measuring the contents of Malondialdehyde (MDA) and Glutathione (GSH) and the activities of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) (Coomassie brilliant blue method).
4.1.4 pathological Observation of liver tissue
Liver tissues of the same parts of each group of experimental mice are taken, washed clean by cold physiological saline and fixed by neutral formalin, paraffin sections with the size of 4 mu m are prepared, HE staining is carried out, and the morphological change of the paraffin sections is observed under a microscope.
4.1.5 Experimental data processing
The experimental data are all represented by mean values plus or minus standard errors (x plus or minus s), and the data are subjected to the homogeneity test of variance; data comparisons between groups were performed using one-way analysis of variance. Significance was analyzed using the Duncan test in one-way analysis of variance (ANOVA) with SPSS19.0 statistical software, with P <0.05 being significantly different and P <0.01 being significantly different.
4.2 results and analysis
4.2.1 Effect of plant functional salts on body weight of laboratory mice
In the test period of the period 21d, the body mass of the mice of each experimental group is prolonged along with the feeding time, no significant difference is shown in the body mass (P is greater than 0.05), the skin and hair of the mice of each group are observed to be smooth, the clinical activity performance is normal, the health condition is good, and the fact that the clinical performance of the mice is not influenced by taking different salt samples is shown. The body mass of each group of experimental mice in the experimental period is detailed in Table 10.
TABLE 10 comparison of body constitution of mice of each experimental group in the test period
Note: if letters in two same columns are completely different, the two columns have obvious difference (P < 0.05);
4.2.2 Effect of plant functional salts on transaminase activity in serum of mice with chemical liver injury
As can be seen from Table 11, model group (3)#) The serum glutamic acid transaminase (ALT) and aspartate transaminase (AST) activities of the mice are both remarkably higher than those of a normal control group (2)#And 1#)(P<0.01), indicating that molding was successful.
As can be seen from Table 11, all three plant functional salts showed varying degrees of activity in reducing serum transaminase, whereas the original salt had little effect. Compared with the model group, the three plant functional salts have extremely obvious effect of inhibiting the activity increase of ALT (P)<0.01); pine salt (P) in inhibiting AST activity increase<0.01) to the remaining two samples (P)<0.05) more pronounced; pine salt pair CCl4The inhibition effect of the acute liver injury mouse serum transaminase sharply rising is better than that of bamboo salt and plum salt. The bamboo salt, the pine salt and the plum salt have certain liver protection effects. However, the positive drug control group (bifendate) had no effect on AST although it had the strongest efficacy in inhibiting ALT activity. The commercially available Renshansa edulis salt does not show the inhibitory effect on the serum transaminase activity of the mice with acute chemical liver injury, like the original salt.
TABLE 11 comparison of transaminase activities in serum of mice of each experimental group
Note: if letters in two same columns are completely different, the two columns have obvious difference (P < 0.05);
model control group (3)#) Control group with NaCl (2)#) In contrast to the above-mentioned results,#represents P<0.01;
Each test group (4)#~9 #) Control group with model (3)#) By comparison, denotes P<0.01。
4.2.3 Effect of plant functional salts on oxidative stress index of mice with chemical liver injury
When an experimental animal is attacked by a chemical poison, an important characterization of acute liver injury is that the MDA content is increased, the GSH level is reduced, the SOD activity is reduced and the like, and strong oxidative stress reaction is generated along with the peroxidation of liver tissue membrane lipid.
The data in Table 12 show that two groups (2) ingested with the same salt-free feed and simultaneously gavage with 4% pure NaCl#And 3#) By CCl4After the model is made, various oxidative stress indexes are obviously changed, and particularly the GSH and MDA levels are obviously changed (P)<0.01), indicating gavage CCl4The liver of the latter animals was indeed damaged.
From table 12, it can be seen that three plant functional salts can increase the GSH content of the damaged liver to some extent, wherein the prunus mume salt shows a very significant enhancing effect; and can reduce MDA content of damaged liver, wherein the strongest action is plum salt (P <0.01), and the second is Renshanzizhu salt and pine salt (P < 0.01); the bifendate group also showed the effects of raising GSH and reducing MDA content, but none was as effective as the prunus salt. In the aspect of improving the activity of endogenous antioxidant enzymes of damaged livers, the three plant functional salts all show a certain effect, wherein the bamboo salt simultaneously shows a remarkable influence on the activity of T-SOD (P is less than 0.05).
TABLE 12 comparison of antioxidant Activity of liver tissue of mice in each experimental group
Note: if letters in two same columns are completely different, the two columns have obvious difference (P < 0.05);
model control group (3)#) Control group with NaCl (2)#) In contrast to the above-mentioned results,#represents P<0.01;
Each test group (4)#~9 #) Control group with model (3)#) By comparison, denotes P<0.01。
4.2.4 histopathological Observation of liver injury and repair in Experimental animals
FIG. 14 is the effect of plant functional salts on liver histopathology in experimental mice (. times.400). 1#And 2#The mouse liver tissue is the liver tissue of a normal mouse, so that the liver cell cords are arranged orderly, the cell boundary is clear, the cell karyotype is normal, and no obvious abnormality exists; 3#Hepatic cell cord disorder in NaCl model groupThe karyotype is obviously changed, and a large amount of cells infiltrate the central vein; 4#The bifendate group has clear liver cell cable arrangement, normal cell nucleus type and a small amount of vacuole degeneration, and shows that the bifendate dripping pill has a treatment effect on acute liver injury; 5#The liver cell cables of the Renshan salt group are arranged clearly, the cell nucleus type is normal, and a small amount of vacuole denaturation and water sample denaturation exist; 6#The hepatic cell cords of the original salt control group are not regularly arranged, the cell karyotype is obviously changed, and a large amount of cells infiltrate the central vein; 7#The liver cell cables of the bamboo salt test group are arranged clearly, the cell boundary is clear, the cell karyotype is normal, and a small amount of vacuole degeneration exists; 8#The plum salt test group has clear liver cell cable arrangement, normal cell nucleus type and a small amount of vacuole degeneration; 9#The loose salt test group has clear liver cell cord arrangement, normal cell karyotype, little vacuolar degeneration and little cell infiltration in central venous vessels. According to the morphological examination result of the liver histopathological section, the bamboo salt, the pine salt and the plum salt are all shown to be applied to CCl4The mouse acute liver injury caused by the drug has a certain protection effect.
4.3 conclusion
China is the first major country of the world with liver diseases, and the number of chronic liver diseases is predicted to reach 4.5 hundred million in 2020. The high-incidence national conditions of liver diseases are largely related to food sources, drinking water, eating habits, catering ways and the like. The edible salt for residents in China mainly comes from sea salt, sun-cured sea salt (raw salt), namely a concentrate of seawater, and all pollutants in the ocean are likely to be reserved in the sea salt, so that certain food safety hazards are brought. The research result shows that the GSH level in the liver of the experimental mouse which intakes the original salt is extremely obviously lower than that of the group which intakes the pure NaCl with the same dosage, which indicates that some harmful factors which are adverse to the liver possibly exist in the original salt, and the oxidative stress of the liver is caused. The final result shows that the effective components absorbed from the bamboo tube by the bamboo salt baked by the traditional method are limited, and the liver protection effect of the bamboo salt baked by the traditional method is not as good as that of the pine, bamboo and plum salt provided by the invention.
5.1 materials, reagents and instruments
Mouse B16 melanoma cells, shanghai cell institute of chinese academy of sciences; RPMI-1640 medium, Gibco, USA; fetal bovine serum, ThermoFisher, USA; trypsin, penicillin-streptomycin solution, Phosphate Buffered Saline (PBS), AR, hangzhou kokai biotechnology limited; levodopa (L-DOPA), tyrosine, tyrosinase (25KU, not less than 500u/mg), AR, Beijing Soilebao technologies, Inc.; tetramethyl azo blue (MTT, dye liquor mass fraction is 5g/L), Beijing Ragen biotechnology, Inc.; TritonX-100, -arbutin, dimethyl sulfoxide (DMSO), AR, Shanghai Allantin reagent, Inc.
Eon microplate reader, BioTek corporation, usa; LDZX-50KBS vertical autoclave, Shanghai Shenan medical instrument factory; CP-ST50A type carbon dioxide incubator, Changsha Changjin science and technology Co., Ltd; SW-CJ-F clean bench, Shanghai Boxun industries, Ltd; CKX41 inverted microscope, OLYMPUS optics industries, japan; BDFACSCaLIBUR flow cytometer, BD company, usa.
5.2 Experimental methods
5.2.1 in vitro tyrosinase inhibition assay
Tyrosine is taken as a substrate for measuring the activity of the tyrosinase monophenolase, L-DOPA is taken as a substrate for measuring the activity of the tyrosinase diphenol, and the method of Huang and the like is referred for carrying out the activity measurement of the tyrosinase. Firstly, preparing original salt, roasted salt, pine salt, bamboo salt and plum salt into salt solutions with the mass fractions of 0.2%, 1%, 5% and 10% by using PBS (phosphate buffer solution), sequentially and accurately absorbing sample solutions with different concentrations, 0.1mol/L PBS solution with the pH value of 6.8 and 1.0mmol/L tyrosine or L-DOPA (tyrosine-aspartate) according to a table, fully and uniformly mixing, putting the mixture into a water bath kettle at 25 ℃ for incubation for 10min, then adding corresponding 870u/mg tyrosinase solution, and measuring the light absorption value of each reaction solution at 475nm after 10 min. The inhibition rate was calculated as follows:
tyrosinase monophenolase/diphenolase inhibition rate/% ([ 1- (A3-a4)/(a1-a2) ] × 100
TABLE 13 reaction solution composition (mL) in vitro tyrosinase activity assay
5.2.2 determination of cell proliferation Rate of B16
The proliferation rate of B16 cells was measured by MTT method using the original salt as a negative control, arbutin and Korean bamboo salt as a positive control. Selecting B16 cells in logarithmic phase, inoculating into 96-well plate, adhering to the wall, removing original culture medium, adding culture medium containing 0.25%, 0.50%, 0.75% and 1% of original salt, pine salt, bamboo salt, plum salt, -arbutin and Korean bamboo salt, and culturing at 37 deg.C with 5% CO2Culturing under the condition for 48 h. The sample was removed 4h before the assay, 20. mu. LMTT (5mg/mL) solution was added to each well, CO was added25% CO at 37 ℃ in an incubator2Incubation was carried out for 4h in the environment, then the medium and MTT were discarded, 150 μ l LDMSO was added to each well to dissolve residual MTT-formazan crystals, shaking for 10min, with cells cultured without added sample as control and PBS as blank. The absorbance at 570nm of each well was measured using a microplate reader. The cell proliferation rate was calculated as follows:
cell proliferation rate/% (OD)570 (experiment)-OD 570 (blank))/(OD 570 (control)-OD 570 (blank))×100
5.2.3 determination of tyrosinase Activity in B16 cells
The tyrosinase activity in B16 cells was measured by the dopa oxidation method. B16 cells are inoculated into a 96-well plate, the original culture medium is discarded after adherence, and the culture medium containing each sample is added for culture for 48 h. Pouring out the culture solution from each group of cells with the action time up, washing the cells for 2 times by using PBS buffer solution, adding 50 mu L of 1% by mass of Triton-X100 aqueous solution into each hole, then quickly placing the cells into a refrigerator at the temperature of 80 ℃ below zero for freezing for 30min, taking out the cells, melting the cells at the temperature of 37 ℃ to completely break the cells, adding 10 mu L L-DOPA solution (10g/L) into each hole, taking the cells cultured without adding samples as a control, using PBS as a blank, reacting the cells at the temperature of 37 ℃ for 2h, and measuring the OD value of each hole at the position of 475nm by using a microplate reader. Intracellular tyrosinase activity was calculated as follows:
intracellular tyrosinase activity/% (OD)475 (experiment)-OD 475 (blank))/(OD 475 (control)-OD 475 (blank)) Cell proliferation Rate X100
5.2.4B 16 cell migration assay
B16 cells were selected in the logarithmic growth phase and the cell concentration was adjusted to 4X 105Adding 2mL of the cells into a 6-well plate, discarding the original culture medium when the bottom of the cells is covered by about 60%, drawing a straight line on the bottom of each well by using a 10-microliter pipette tip (one-time operation), slightly washing off the slipped cells by using PBS (phosphate buffer solution), continuously culturing for 48 hours by using a culture medium prepared from 0.5% of original salt, pine salt, bamboo salt, plum salt and arbutin by mass fraction, observing the growth state and migration condition of the cells under a microscope, and taking the cells cultured without the added samples as a control.
5.2.5B 16 cell cycle determination
After B16 cells were allowed to adhere to the walls of the 6-well plates, a medium containing 0.5% of each sample was added for culture, and each group of three was cultured in parallel with a blank control group for 48 hours. After the culture was completed, the cells were collected, washed with PBS, the supernatant was removed, 500. mu.L of a cold 70% ethanol solution was added, fixed overnight, and then centrifuged and the remaining ethanol was aspirated. Adding RnaseA solution of 100 mu L into the cell sediment, resuspending the cells, carrying out water bath at 37 ℃ for 30min, adding 400 mu LPI staining solution, mixing uniformly, incubating at 4 ℃ in the dark for 30min, and transferring to a flow tube for machine-loading detection.
5.2.6 Experimental data processing
Experimental data were processed using Origin9 and calculated results expressed as Mean ± standard deviation (Mean ± SD) and tested for significance using Duncan in SPSS 21 one-way analysis of variance ANOVA.
5.3 analysis of results
5.3.1 inhibitory Activity of plant salts on tyrosinase in vitro
When tyrosine is used as a substrate, tyrosinase can catalyze dopa and further generate melanin, and the process mainly exerts tyrosinase monophenol enzyme activity. The inhibition of tyrosinase monophenolase activity by various salt solutions is shown in figure 15. As can be seen from FIG. 15, the original salt showed no significant inhibition of tyrosinase in the concentration range of 0.2-10.0%. The influence of the roasted salt on tyrosinase is the largest at a concentration of 5.0%, but the inhibition rate is not higher than 10%. The pine salt and the bamboo salt have obvious influence on the activity of tyrosinase and are in a concentration effect relationship, when the mass fraction is 0.2-5.0%, the inhibition rate on the tyrosinase is higher along with the increase of the salt concentration, the inhibition rate reaches more than 94% at 5.0%, and the inhibition rate is basically unchanged at 10.0%. Compared with pine salt and bamboo salt, the plum salt has lower inhibition rate on tyrosinase at low concentration (0.2% and 1%), the inhibition rate is rapidly increased when the concentration is more than 5.0%, the inhibition rate on tyrosinase is 73.2% by 10.0% of plum salt solution, and the effect is slightly inferior to that of pine salt and bamboo salt solution with the same concentration.
When dopa is used as a substrate, tyrosinase can catalyze dopa to generate dopaquinone and further generate melanin, and the process mainly exerts tyrosinase diphenolase activity. The inhibition rate of tyrosinase diphenolase by various salt solutions is shown in FIG. 16. As can be seen from the figure, compared with the inhibition rate of tyrosinase monophenolase, the inhibition rates of raw salt and roasted salt on tyrosinase diphenolase are obviously enhanced, and a certain concentration effect relationship exists, but the maximum inhibition rate is not more than 30%. The tyrosinase diphenolase inhibition effect of pine salt and bamboo salt is similar to that of monophenolase inhibition effect, the diphenolase inhibition rate of high-concentration (5.0% and 10.0%) plum salt solution is not much different from that of pine salt and bamboo salt solution with the same concentration, and the inhibition rate can reach 92.0% when the concentration is 5%. Compared with the original salt, the plant salt has more contents of metal elements such as potassium, iron, magnesium, calcium and the like, and may have certain influence on the protein structure of tyrosinase, so that the activity of the tyrosinase is inhibited.
5.3.2 Effect of plant functional salts on B16 cell proliferation
Note: different letters indicate significant differences at the same concentration (p <0.05) and n 6.
Melanin is produced by melanocytes, and generally, inhibition of melanocyte proliferation reduces melanin production. The proliferation rates of mouse B16 melanocytes in samples of different concentrations are shown in table 14. As can be seen from table 14, the salt solutions with concentrations of 0.25% or more inhibited the growth of B16 cells, and the proliferation rates of the cells decreased with increasing salt concentrations, and at 0.75%, the proliferation rates of the cells in the salt groups were not much different, and the growth of the cells was severely affected by the salt solution with concentration of 1%, and the proliferation rates were all less than 10%. In the low concentration range (0.25%, 0.5%), the proliferation rate of B16 cells in pine salt, bamboo salt, plum salt and Korean bamboo salt is not lower than that of original salt, so compared with the original salt, the plant salt has no obvious additional cytotoxicity to B16 cells, and the proliferation rate of cells in 0.5% of pine salt, bamboo salt and plum salt is far higher than that of the original salt, which indicates that the plant salts have a certain protection effect on B16 cells in NaCl environment. The positive control arbutin group has a small relationship between cell proliferation rate and concentration, and the cell proliferation rate and the concentration are both about 70%.
5.3.3 Effect of plant salts on tyrosinase Activity in B16 cells
Tyrosinase is a key rate-limiting enzyme for synthesizing melanin by B16 cells, and the enzyme activity of the tyrosinase is an important influencing factor for melanin generation and an important index for measuring the whitening effect of a functional factor. As can be seen from table 15, each sample had a certain effect on the tyrosinase activity of B16 cells, and the intracellular tyrosinase activity was significantly decreased, and was in a certain concentration effect relationship. Compared with the original salt, the cell tyrosinase activities of the pine salt, the bamboo salt and the Korean bamboo salt are higher in each group, which shows that the plant salts have no inhibiting effect on B16 cell tyrosinase as compared with the original salt, and the cell tyrosinase activity of the prunus mume salt is smaller than that of the original salt under each concentration, which shows that the prunus mume salt can obviously reduce the tyrosinase activity of B16 cells and has an effect superior to that of a test object in each group. The cell tyrosinase activities of the positive control arbutin group are all higher than those of the salt sample group with the same concentration.
Note: different letters indicate significant differences at the same concentration (p <0.05) and n 6.
5.3.4 Effect of plant salts on migration of B16 cells
The important characteristic of tumor cells is that they can diffuse and metastasize during proliferation, and their ability to diffuse can be judged by their mobility. As can be seen from FIG. 17, the streaked area of the B16 cells in the normal group (a) was substantially healed after 48h of culture, indicating that the cells had a strong migration ability. When the mass fraction of the test substance is 0.5%, only part of the cells of B16 acted by the original salt (B) enter the marked area, which indicates that the original salt has a certain inhibition effect on cell migration. In cells acted by the pine salt (d), the bamboo salt (e) and the plum salt (f), the scribed area basically has no cell entering, which shows that the three plant salts can obviously inhibit the transfer of the B16 cells and have stronger effect than the original salt. Arbutin (c) also has some inhibitory effect on the mobility of B16 cells, but its effect is not as good as that of plant salts.
5.3.5 Effect of plant salts on B16 cell cycle
The cell cycle refers to the process from the end of cell division to the end of the next cell division, and is divided into four phases, the G0/G1 phase (pre-DNA synthesis phase), the S phase (DNA replication phase), the G2 phase (DNA replication completion to mitosis initiation), and the M phase (cell division initiation to end). The DNA content in the cells in different periods is different, the cell DNA is dyed by using the fluorescent dye PI, and the fluorescence intensity is detected in a flow cytometer, so that the distribution of the cells in different periods can be researched, and the proliferation condition of the cells can be known. As can be seen from Table 16, the distribution of the original salt and arbutin in the G0/G1 phase and the S phase of the normal group cells is not significantly different, while the distribution ratio of the pine salt, bamboo salt and plum salt in the S phase and the G2/M phase is significantly increased, and the distribution in the G0/G1 phase is less. The sum of the S phase and G2/M represents the proportion of the cells in the process of DNA replication and mitosis, and can represent the proliferation capacity of the cells, and the results show that the proliferation capacities of the cells in the pine salt group and the bamboo salt group are better, the proliferation capacities of the other groups are second, and the proliferation capacities of the other groups are not obviously changed. The MTT results showed that each test substance had a certain inhibitory effect on the proliferation of B16 cell population, whereas the cell cycle studies found that the proliferation ability of the surviving cells was good, and it was presumed that the treatment with the test substance might cause the death of some cells, and that it had the effect of promoting the proliferation of some of the tolerant cells.
TABLE 160.5% Mass fraction Effect of each sample on B16 cell cycle
Note: different letters indicate significant differences (p <0.05) at the same cell cycle, with n-3.
5.4 conclusion
The pine, bamboo and plum salt provided by the invention is taken as three representative plant functional salts, shows concentration-dependent inhibition effects on monophenolase activity and diphenolase activity of tyrosinase in vitro, and when the concentration of a sample is 10%, the inhibition rates of the tyrosinase monophenolase respectively reach 97.8%, 94.2% and 73.3%; when the concentration is 5%, the inhibition rates of the diphenolase reach 99.9%, 93.0% and 92.0% respectively. The plant functional salt has certain inhibition effect on B16 cell proliferation, but has no obvious cytotoxicity compared with the original salt. The tyrosinase activity in B16 cells can be obviously reduced by the plum salt, and the tyrosinase activity in B16 cells in a 0.75% plum salt culture medium is only 6.07%. Meanwhile, the plant functional salt can remarkably inhibit the lateral migration of the B16 cell, but does not inhibit the proliferation of the B16 cell by influencing the cell cycle. The plant functional salt is used as an in vitro and intracellular tyrosinase inorganic inhibitor, has good inhibition effect, and can be considered as a fruit and vegetable preservative and a whitening functional component for human bodies.
6.1 sources and groups of Experimental animals
Experimental animals: the male rat SD rat is 13-14 weeks old and has a body weight of about 200 g. One week after adaptive feeding, the animals were randomly divided into 8 groups, namely, a normal saline feed control group, a gavage normal saline non-saline feed control group, a high-salt model group, an experimental group (pine, bamboo and plum plant functional salts) and a positive control group (felodipine sustained-release tablets), wherein 4 animals were fed in each group, and the experimental design and dosage configuration are shown in table 17. During the experiment, the animals are raised in a single cage, the raising temperature is 25 ℃, the humidity is 30 percent, and the clinical performance is observed every week. In the administration stage, the daily high salt intake of the rat is 4000mg/kg (equivalent to 8% of salt content in daily ration), each sample and the control sample are prepared into solutions with corresponding concentrations under the premise of freely drinking purified water, and the rat is orally fed with 2mL/100g of bamboo saline, pine saline, plum saline and the like every day. Except for normal control group (1)#) Except for the conventional feed, the rest 8 groups all use the salt-free basic feed.
TABLE 17 animal test groups and dosages administered
To ensure comparability of the test data, free diet conventional feed containing 0.3% common salt (group 1)#) And a control group (group 2) fed with the same quality of the original saline solution through oral feeding and simultaneously fed with the salt-free feed#) The effect of salt intake on rats in the gavage mode was compared to that in the feed.
High salt feed group addition amount 8% salt was added to the reference feeds converted to gavage concentration.
6.2 test conditions
Blood pressure and body weight were measured every 4 days for 27 days. After the last gastric lavage, fasting is carried out without water prohibition, and the gastric lavage liquid is placed in a metabolism cage for 24 hours and urine is collected. After the end of the abdominal injection, the rats were sacrificed after the heart was bled.
6.3 test results
6.3.1 weight changes in Experimental rats
The weights of animals in each group have no obvious difference within 24 days of the experiment, and the weight is not obviously influenced by gastric lavage and high salt intake.
TABLE 18 weight change table
Compared with the conventional raw salt for intragastric administration (group 2),Δrepresents p<0.05, ΔΔRepresents p<0.01;
Compared with the normal group (group 2) of the original salt,#represents p<0.05, ##Represents p<0.01;
Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8),*represents p<0.05, **Represents p<0.01。
6.3.2 changes in systolic blood pressure in Experimental rats
The significant difference between the high-salt experimental group 3 and the conventional original salt group 2 occurs by the time of intragastric administration of the high-dose salt to the 20 th day, which shows that hypertension is caused by oral administration of the high-dose salt, and the experimental modeling is successful. From the experimental group, the plant functional salt groups of pine, bamboo and plum obtained in the examples all have significant differences from the original salt group by the 24 th day, which shows that the plant functional salt prepared in the example has the effect of preventing hypertension by eating the plant functional salt. It is noted that the blood pressure lowering effect of the plum salt in the plant salt is most obvious, and has no significant difference with low salt intake, and is possibly related to the special physical and chemical structure and element content of the plum salt.
TABLE 19 blood pressure variation values of rats
Compared with the conventional raw salt for intragastric administration (group 2),△represents p<0.05, △△Represents p<0.01;
Compared with the normal group (group 2) of the original salt,#represents p<0.05, ##Represents p<0.01;
Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8),*represents p<0.05, **Represents p<0.01。
6.3.3 urine volume Change in Experimental rats over 24h
The conventional feed and the conventional raw salt group have no significant difference, which shows that the feeding has no obvious influence on the urine volume. The urine volume was significantly higher in the high salt group than in the normal salt group due to increased water intake resulting from ingestion of high concentrations of saline. However, in the plant functional salt, the urine volume of the bamboo salt is obviously higher than that of other groups, and the obvious difference is formed between the bamboo salt and the original salt group, which indicates that the bamboo salt has the obvious diuretic effect. It is noted that the urine volume of the bamboo salt group is higher than that of the positive control, and felodipine is a selective calcium ion antagonist, mainly inhibits the inflow of calcium outside arteriolar smooth muscle cells, and has the functions of promoting natriuresis and diuresis. The results show that the diuretic effect of the bamboo salt is particularly remarkable.
TABLE 20 urine volume values of rats
Compared with the conventional raw salt for intragastric administration (group 2),△represents p<0.05, △△Represents p<0.01;
Compared with the normal group (group 2) of the original salt,#represents p<0.05, ##Represents p<0.01;
Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8),*to representp<0.05, **Represents p<0.01。
6.3.4 analysis result of serum biochemical index of experimental rat
(1) Liver function
Alanine Aminotransferase (ALT), an enzyme involved in protein metabolism in humans, is an important indicator of the degree of liver disease. ALT can accelerate the transformation of protein amino acid in vivo, and the content of ALT is the maximum in liver cells. When the tissue organ is diseased, ALT in the tissue organ is released into blood, so that the ALT content in the serum is increased. Compared with normal rats, ALT in serum of the rats in the high-salt group is remarkably increased, which indicates that the liver is damaged to some extent.
Aspartate Aminotransferase (AST) is helpful in understanding the extent of damage to heart muscle, liver, and kidney tissues. AST is present in various tissues of the human body, with the most abundant content of cardiac muscle, followed by the liver. When necrosis of heart and liver cells occurs, m-AST is released from mitochondria, resulting in increased AST in serum. The AST in the serum of the rat in the high-salt group is obviously increased, which indicates that the liver is influenced after high-concentration salt is ingested.
Alkaline phosphatase (ALP) is widely distributed in human liver, bone, intestine, kidney, placenta and other tissues, is discharged out of the gallbladder through the liver, is mainly used for diagnosis and differential diagnosis of bone, liver and gallbladder system diseases, and the pathological increase of ALP is closely related to the bone, liver and gallbladder diseases. There was no significant difference in ALP content among the groups, indicating that ingestion of a high salt diet within one month had a less pronounced effect on ALP.
TABLE 21 liver function indices
Compared with the conventional raw salt for intragastric administration (group 2),△to representp<0.05, △△Represents p<0.01;
Compared with the normal group (group 2) of the original salt,#represents p<0.05, ##Represents p<0.01;
Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8),*represents p<0.05, **Represents p<0.01。
(2) Renal function
Urea Nitrogen (BUN) is a nitrogen-containing compound in plasma other than proteins, and is filtered from the glomerulus and excreted. Upon decompensation of renal insufficiency, BUN will rise. Clinically using the glomerular filtration function as an index for judging the glomerular filtration function; creatinine (CRea) is a product of in vivo metabolism of muscle tissues, is mainly filtered by glomeruli and is completely discharged out of the body, and when the kidney function is incomplete, the creatinine can be accumulated in the body to become a toxin harmful to the human body; when all uric acid in blood is filtered from glomeruli, abnormal renal handling of urine results in increased levels of Uric Acid (UA) in blood. It can be seen from the table that there is no significant difference in the index of each group within one month, and it is likely that the high-salt diet time is too short.
TABLE 22 renal function indices
Compared with the conventional raw salt for intragastric administration (group 2),△represents p<0.05, △△Represents p<0.01;
Compared with the normal group (group 2) of the original salt,#represents p<0.05, ##Represents p<0.01;
Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8),*represents p<0.05, **Represents p<0.01。
(3) Myocardial function
Skeletal muscle, cardiac muscle and smooth muscle contain a large amount of Creatine Kinase (CK), and are mainly present in cytoplasm and mitochondria, and are important kinases directly related to intracellular energy transfer, muscle contraction and ATP regeneration. The creatine kinase activity assay can be used for diagnosis of skeletal muscle diseases and cardiac muscle diseases. Pathological increase: myocardial infarction, viral myocarditis, dermatomyositis, muscular dystrophy, pericarditis, cerebrovascular accident rupture, etc. It can be seen from the table that ingestion of high concentrations of salt has a significant effect on CK levels, and that a high salt diet more than one month may have some effect on myocardial function. However, it is noted that the CK content of the pine salt is significantly lower than that of the high-salt group, which indicates that the pine salt has potential in preventing myocardial damage.
Lactate Dehydrogenase (LDH) is present in almost all tissues, with heart, skeletal muscle and kidney being the most abundant and useful for the diagnosis of myocardial diseases. Lactate dehydrogenase increases: mainly seen in myocardial infarction, hepatitis, malignant tumor, pulmonary infarction, leukemia, hemolytic anemia, kidney diseases, progressive muscular atrophy and the like. The high-salt diet had lower CK and LDH contents than the normal salt diet group, further illustrating the impairment of myocardial function by the high-salt diet.
TABLE 23 myocardial function indices
Compared with the conventional raw salt for intragastric administration (group 2),△represents p<0.05, △△Represents p<0.01;
Compared with the normal group (group 2) of the original salt,#represents p<0.05, ##Represents p<0.01;
Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8),*represents p<0.05, **Represents p<0.01。
(4) Serum ion content
The experiment result of one month shows that the concentration difference of each ion in the serum is not large.
TABLE 24 serum ion content
Compared with the conventional raw salt for intragastric administration (group 2),△represents p<0.05, △△Represents p<0.01;
Compared with the normal group (group 2) of the original salt,#represents p<0.05, ##Represents p<0.01;
Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8),*represents p<0.05, **Represents p<0.01。
6.3.5 urine index analysis results of Experimental rats
(1) Total excretion
Urinary creatinine (CRea) is derived primarily from blood, and is removed from the body via glomerular filtration with little absorption and excretion by the renal tubules. Urinary creatinine levels change when there is a problem with the kidneys. Normal urine contains a small amount of small molecular protein (U-TP), and proteinuria is formed when protein in urine is increased, which is a common manifestation of kidney diseases, and also can occur in systemic diseases. Urine microalbumin (U-ALB) is one of the important plasma proteins, and under normal conditions, albumin has a large molecular weight and cannot cross the glomerular basement membrane. In the disease, glomerular basement membrane is damaged to change permeability, so that albumin is excreted into urine, and the urine albumin concentration is continuously increased. From the table, it can be seen that the high-salt diet has significant difference on the U-TP and U-ALB contents of SD rats in one month, which indicates that the kidney function is damaged, but the serum kidney index and the CRea content in urine have no significant change, which indicates that the high-salt diet has not strong influence degree in one month, and if the high-salt diet is used for a long time, the kidney is damaged. It is noted that the loose salt U-TP level was significantly lower than the high salt group, comparable to the positive control, indicating that loose salt also had the effect of protecting renal function.
TABLE 25 Biochemical indicators of urine
Compared with the conventional raw salt for intragastric administration (group 2),△represents p<0.05, △△Represents p<0.01;
Compared with the normal group (group 2) of the original salt,#represents p<0.05, ##Represents p<0.01;
Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8),*represents p<0.05, **Represents p<0.01。
(2) Excretion of calcium, phosphorus, magnesium, sodium, chloride, potassium ions in urine
Besides the increase of Na and Cl ion content, the contents of K, Ca and P in the high-salt group are obviously increased, wherein the P element in the bamboo salt is obvious. In the plant salt element analysis, neither the original salt nor the roasted salt P element is detected, and the salt of the pinus sylvatus is detected to be 71.8, 45.3 and 424mg/kg respectively. The physical and chemical indexes of the plant functional salt show that the element content is obviously higher than that of the original salt and the roasted salt, but the urine excretion amount has no obvious difference, which indicates that the rich elements in the plant functional salt are absorbed by the body.
TABLE 26 urinary Ionic excretion
Compared with the conventional raw salt for intragastric administration (group 2),△represents p<0.05, △△Represents p<0.01;
High salt test group (group 3) Normal to Normal saltIn comparison with the group (group 2),#represents p<0.05, ##Represents p<0.01;
Compared with the high-salt test group (group 3), each of the high-salt test groups (groups 4 to 8),*represents p<0.05, **Represents p<0.01。
6.4 conclusion
The common salt and the roasted salt have no significant difference in the test for preventing the hypertension of SD rats, which shows that the roasted salt without the added plant extract prepared by the invention has no special biological efficacy.
After a month of high-salt diet, the systolic pressure of male SD rats is obviously increased after the male SD rats eat the raw salt, and obvious liver, kidney and myocardial function damage is shown; the blood pressure of the experimental animals which ingest the plant functional salt is obviously lower than that of the original salt, which shows that the plant functional salt has the potential of replacing common edible salt and preventing and treating hypertension.
All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.
Claims (13)
- A functional plant salt, which is prepared by the following steps: one or more plant extract powders are evenly mixed with fine salt according to the addition amount of 1-40 percent and baked at the high temperature of 800-1400 ℃ for one time or several times to obtain the functional plant salt.
- The functional plant salt of claim 1, wherein the plant extract is an extract of a substance selected from the group consisting of: plum, pine, tea, lotus, chrysanthemum, pomegranate, Chinese flowering crabapple, edible fungus and algae.
- The functional plant salt of claim 1, wherein the plant extract is a plum extract and/or a pine extract.
- The functional plant salt of claim 3, wherein the plum extract is selected from the group consisting of: plum extract, plum leaf extract, plum branch extract, plum root extract, or a combination thereof; and/orThe pine extract is selected from the group consisting of: pine needle extract, pine bark extract, pine pollen extract, or a combination thereof.
- The functional plant salt of claim 1, wherein the fine salt is selected from the group consisting of: sea salt, lake salt, well salt and mineral salt.
- The functional plant salt of claim 1, wherein the fine salt is crude sea salt.
- The functional plant salt according to claim 1, wherein the functional plant salt is pine salt and/or plum salt.
- The functional plant salt of claim 1, wherein the plant extract further comprises a bamboo extract.
- A method for preparing the functional plant salt according to claim 1, comprising the steps of: one or more plant extract powders are evenly mixed with fine salt according to the addition amount of 1-40 percent and baked at the high temperature of 800-1400 ℃ for one time or several times to obtain the functional plant salt.
- The method of claim 9, wherein the cooking is performed in a mechanized oven.
- The method of claim 9, wherein the roasting is performed in a medium frequency furnace.
- A food or pharmaceutical composition comprising one or more functional plant salts of claim 1.
- Use of the functional plant salt of claim 1 for the preparation of a substance selected from the group consisting of: pharmaceutical adjuvants, health food/beverage, functional food/beverage, table salt, cooking salt, special-purpose cosmetics or personal care products.
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CN111543619A (en) * | 2020-05-25 | 2020-08-18 | 任玉龙 | Preparation method of selenium bamboo salt |
CN112586718B (en) * | 2020-12-18 | 2022-12-02 | 湖北省益欣盐产业技术研究院有限公司 | Selenium-rich edible salt and preparation method thereof |
CN112683632A (en) * | 2021-01-13 | 2021-04-20 | 贵州省农作物品种资源研究所 | Method for determining content of bletilla polysaccharide in bletilla striata tubers |
CN113768125A (en) * | 2021-07-27 | 2021-12-10 | 浙江竹研科技有限公司 | Preparation method of bamboo salt containing peptides and bamboo salt containing peptides prepared by preparation method |
CN116158524A (en) * | 2022-12-22 | 2023-05-26 | 唐山市银海食盐有限公司 | Preparation method of blue bamboo salt |
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